CN114652717B - Pharmaceutical application of naphazoline hydrochloride - Google Patents
Pharmaceutical application of naphazoline hydrochloride Download PDFInfo
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- CN114652717B CN114652717B CN202011525316.4A CN202011525316A CN114652717B CN 114652717 B CN114652717 B CN 114652717B CN 202011525316 A CN202011525316 A CN 202011525316A CN 114652717 B CN114652717 B CN 114652717B
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- naphazoline
- cardiomyocytes
- pharmaceutically acceptable
- acceptable salt
- proliferation
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- 229960004760 naphazoline hydrochloride Drugs 0.000 title description 12
- CNIIGCLFLJGOGP-UHFFFAOYSA-N 2-(1-naphthalenylmethyl)-4,5-dihydro-1H-imidazole Chemical compound C=1C=CC2=CC=CC=C2C=1CC1=NCCN1 CNIIGCLFLJGOGP-UHFFFAOYSA-N 0.000 claims abstract description 30
- 229960005016 naphazoline Drugs 0.000 claims abstract description 25
- 150000003839 salts Chemical class 0.000 claims abstract description 18
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- AIYUHDOJVYHVIT-UHFFFAOYSA-M caesium chloride Chemical compound [Cl-].[Cs+] AIYUHDOJVYHVIT-UHFFFAOYSA-M 0.000 description 1
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- BFMYDTVEBKDAKJ-UHFFFAOYSA-L disodium;(2',7'-dibromo-3',6'-dioxido-3-oxospiro[2-benzofuran-1,9'-xanthene]-4'-yl)mercury;hydrate Chemical compound O.[Na+].[Na+].O1C(=O)C2=CC=CC=C2C21C1=CC(Br)=C([O-])C([Hg])=C1OC1=C2C=C(Br)C([O-])=C1 BFMYDTVEBKDAKJ-UHFFFAOYSA-L 0.000 description 1
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- 210000000130 stem cell Anatomy 0.000 description 1
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- 230000000699 topical effect Effects 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- 241000701161 unidentified adenovirus Species 0.000 description 1
- 239000002550 vasoactive agent Substances 0.000 description 1
- 230000009385 viral infection Effects 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- 239000000080 wetting agent Substances 0.000 description 1
Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
- A61K31/4164—1,3-Diazoles
- A61K31/4174—Arylalkylimidazoles, e.g. oxymetazolin, naphazoline, miconazole
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/04—Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0657—Cardiomyocytes; Heart cells
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/80—Neurotransmitters; Neurohormones
- C12N2501/81—Adrenaline
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- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
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- General Chemical & Material Sciences (AREA)
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- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Zoology (AREA)
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- Genetics & Genomics (AREA)
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- Cell Biology (AREA)
- Vascular Medicine (AREA)
- Microbiology (AREA)
- Urology & Nephrology (AREA)
- Hospice & Palliative Care (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a pharmaceutical application of naphazoline. The application is the application of naphazoline or pharmaceutically acceptable salt thereof in preparing medicaments for promoting myocardial cell proliferation. Experiments prove that the naphazoline can promote the proliferation of rat myocardial cells. Therefore, the naphazoline can be used for preparing medicaments for promoting myocardial cell proliferation, medicaments for treating or preventing heart diseases and other related fields, and provides a new medicament and treatment idea for treating or preventing heart diseases such as myocardial infarction.
Description
Technical Field
The invention belongs to the field of medicines, and particularly relates to a pharmaceutical application of naphazoline hydrochloride.
Background
Cardiovascular disease has become the first killer threatening human health, and only heart failure patients are 4000 tens of thousands worldwide, and have become the leading cause of human death. It was found that after adult mammalian cardiomyocytes gradually lose proliferation capacity, and once myocardial infarction occurs, the loss of cardiomyocytes is irreversible. About 20-40 million cardiac myocytes in adults can cause about 25% of cardiac myocytes to be lost within hours after myocardial infarction occurs, and the remaining cardiac myocytes have very limited proliferation capacity and are insufficient to restore contractile function of the heart, and the patient eventually dies. To solve this problem, basic transformation studies have mainly involved searching for cardiac stem cells or precursor cells, transplanting them to the myocardial infarction area by inducing differentiation, but lack of sufficiently authentic molecular markers, low transplanting efficiency, limited the use of this technique; the use of reprogramming techniques to transdifferentiate fibroblasts into cardiomyocytes or to supplement lost myocardium by promoting proliferation of endogenous cardiomyocytes. Previous studies have shown that lower vertebrates such as zebra fish, salamander, etc. have a strong ability to regenerate their hearts, and that neonatal rats also have a certain ability to regenerate, mainly through injury-induced cardiomyocyte proliferation, which is lost after adulthood. Adult hearts have previously been considered unable to regenerate, but there is a great deal of evidence that mammalian cardiomyocytes are slowly updated, and studies have found that neonatal cardiomyocytes are derived from existing myocardium. Therefore, more and more scientists are interested in a strategy that induces proliferation of endogenous cardiomyocytes. The search for drugs that induce endogenous cardiomyocyte proliferation is therefore of great interest for the treatment of cardiovascular diseases in humans.
Naphazoline hydrochloride (Naphazoline Hcl) is a vasoactive drug acting on the circulatory system, belongs to an adrenomimetic drug, has pharmacological activity of constricting blood vessels, and relieves swelling and congestion of nasal mucosa. It is a direct acting alpha adrenergic agonist, which contracts the arterioles dilated intranasally, but has no effect on the beta adrenergic receptor. After topical application, the effect appears within 10 minutes for 2-6 hours. It can be used for treating common cold allergic rhinitis, inflammatory nasal congestion, and acute and chronic rhinitis. However, there is no report on naphazoline hydrochloride in inducing endogenous cardiomyocyte proliferation.
Disclosure of Invention
The invention aims to provide a new application of naphazoline or a pharmaceutically acceptable salt thereof.
The structural formula of the naphazoline is shown as formula I, wherein the naphazoline is CAS No. 550-99-2:
the pharmaceutically acceptable salt may specifically be a hydrochloride salt.
The novel application of naphazoline or the pharmaceutically acceptable salt thereof provided by the invention is the application of naphazoline or the pharmaceutically acceptable salt thereof in preparing products for promoting myocardial cell proliferation.
The cardiomyocytes can be human or mammalian cardiomyocytes. The product may be a pharmaceutical product.
It is another object of the present invention to provide the use of naphazoline or a pharmaceutically acceptable salt thereof for the preparation of a product for the prevention and/or treatment of cardiovascular diseases. The product may be a pharmaceutical product.
Further, in some embodiments of the invention, the cardiovascular disease may be a heart disease caused by loss, injury or death of cardiomyocytes.
Further, in some embodiments of the invention, the heart diseases described above include, but are not limited to, myocardial infarction, heart failure, and other cardiomyopathy resulting from loss, injury, or death of cardiomyocytes, and the like.
Products for promoting myocardial cell proliferation prepared by taking naphazoline or pharmaceutically acceptable salts thereof as active ingredients and products for preventing and/or treating cardiovascular diseases prepared by the naphazoline or pharmaceutically acceptable salts thereof also belong to the protection scope of the invention.
If necessary, one or more pharmaceutically acceptable carriers can be added into the medicine; the carrier includes diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption promoters, surfactants, adsorption carriers, lubricants, etc. which are conventional in the pharmaceutical field.
The medicine can be prepared into various forms such as injection, tablet, powder, granule, capsule, oral liquid, ointment, cream, etc.; the medicaments of the various formulations can be prepared according to the conventional method in the pharmaceutical field.
The above medicine can be introduced into organism such as muscle, intradermal, subcutaneous, intravenous, and mucosal tissue by injection, nasal drop, eye drop, permeation, absorption, physical or chemical mediation; or mixed or wrapped with other substances and introduced into the body.
It is still another object of the present invention to provide a method for culturing cardiomyocytes in vitro.
The method for culturing myocardial cells in vitro provided by the invention comprises the step of adding naphazoline or pharmaceutically acceptable salt thereof into a culture medium containing myocardial cells.
The final concentration of the naphazoline or the pharmaceutically acceptable salt thereof in the culture medium is 0.5-2 mu mol/L. The cardiomyocytes can be human or mammalian cardiomyocytes.
The invention also provides a method for promoting myocardial cell proliferation in vitro.
The method for promoting myocardial cell proliferation in vitro provided by the invention comprises the step of treating myocardial cells with naphazoline or pharmaceutically acceptable salt thereof.
The concentration of the naphazoline or the pharmaceutically acceptable salt thereof in the treatment system is 0.5-2 mu mol/L. The cardiomyocytes can be human or mammalian cardiomyocytes.
Experiments prove that the naphazoline or the pharmaceutically acceptable salt thereof can promote the proliferation of rat myocardial cells. Therefore, the naphazoline or the pharmaceutically acceptable salt thereof can be used for preparing medicaments for promoting myocardial cell proliferation, medicaments for treating or preventing heart diseases and other related fields, and provides a new medicament and treatment idea for treating or preventing heart diseases such as myocardial infarction.
Drawings
FIG. 1 is a graph showing the effect of naphazoline hydrochloride on promoting proliferation of myocardial cells in neonatal rats.
Detailed Description
The invention will be further illustrated with reference to the following specific examples, but the invention is not limited to the following examples. The methods are conventional methods unless otherwise specified. The starting materials are available from published commercial sources unless otherwise specified.
The experimental methods used in the following examples are conventional methods unless otherwise specified.
Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.
Naphazoline hydrochloride used in the following examples was a DMSO solution of naphazoline hydrochloride. Naphazoline HCl manufacturer TargetMol, cat# T6645.
In the following examples, "FBS" is fetal bovine serum.
The construction method of cTnT-mAG-hGmminin (1/110) virus used in the following examples is as follows:
the cTnT myocardial specificity promoter (shown as sequence 1 in a sequence table) and mAG-hGeminin (1/110) (GenBank: NM_ 015895) are assembled and cloned on a pShuttle vector (Addgene 16402) through pEASY-Uni Seamless Cloning and Assembly Kit (full golden CU 101-01), and then restriction enzyme PmeI is adopted for enzyme digestion, and linearization plasmids are collected; co-transformation of the linearized plasmid with pAdEasy1 DNA (Addgene 16400) into BJ5183 competence, selection of recombinant plasmid and amplification followed by transfection of 293A cells with the recombinant plasmid (7.5X10 day before transfection) 5 The 293A cells were plated in 60mm dishes and cultured in DMEM medium containing 5% fetal calf serum until the number of cells reached 1.0-1.5X10 6 Then, the recombinant plasmid is transfected into cells by a calcium phosphate coprecipitation method, the culture solution containing coprecipitation particles is removed the next day after transfection, PBS buffer solution is used for cleaning, then the cells are divided into 6-well plates (3 ml of DMEM culture solution containing 5% fetal bovine serum is added into each well), and the cells are kept stand for 6 hours to adhere to the walls; after 6 hours agarose was covered for the formation of viral plaques (plaques should form within 10-21 days, with the addition of agarose/DMEM mixtures every 4-5 days or when the medium turns yellow); after the initial virus is obtained, the adenovirus is purified by 2-3 rounds of amplification and cesium chloride density gradient centrifugation.
Example 1 in vitro test of Naphazoline hydrochloride (Naphazoline Hcl) to promote proliferation of rat cardiomyocytes
(1) Culture of cardiomyocytes in SD rats
Cardiomyocytes from SD rats at birth were isolated and cultured in DMEM high glucose medium (Hyclone) +5% horse serum (GIBCO) at 37℃in a 5% carbon dioxide incubator.
(2) Experimental grouping and processing
Cardiomyocytes from SD rats were isolated, treated with 5% horse serum (GIBCO) +DMEM high-sugar medium (Hyclone) and cytarabine (final concentration 20 umol/L) to inhibit growth of non-cardiomyocytes, infected with cTnT-mAG-hGeminin (1/110) virus (MOI value of virus infection=100) after 48h of cell attachment, and changed to DMEM medium containing 0.5% FBS after 24 hours, and treated by group dosing as follows:
a. experimental group: naphazoline hydrochloride (final concentration in medium 1. Mu. Mol/L) was added for 24 hours.
b. Blank control group: equivalent DMSO treatment as in experimental group a was added.
(3) Test method
After 24h of treatment of the above 2 groups, nuclei were stained with hoechst fluorochrome, and then photographed with a high content living cell analyzer (Molecular Device), and analyzed for mAG-hGeminin (1/110) -positive cardiomyocyte and total cell count.
(4) Results
The test results are shown in FIG. 1. As can be seen from FIG. 1, mAG-hGeminin (1/110) -positive cardiomyocytes were increased by 2.7-fold compared to the blank control (0.5% FBS+DMSO) after naphazoline hydrochloride treatment. Mean±sem; * P < 0.005.
SEQUENCE LISTING
<110> Xin you kang medicine technology (Nanjing) Nanjing Jing Ruikang molecular medicine technology Co., ltd
New use of naphazoline hydrochloride medicine
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 836
<212> DNA
<213> Artificial sequence
<400> 1
tgtagttaat gattaacccg ccatgctact tatctaccag ggtaatgggg atcctctaga 60
actatagcta gaattcgccc ttacgggccc cccctcgagg tcgggataaa agcagtctgg 120
gctttcacat gacagcatct ggggctgcgg cagagggtcg ggtccgaagc gctgccttat 180
cagcgtcccc agccctggga ggtgacagct ggctggcttg tgtcagcccc tcgggcactc 240
acgtatctcc gtccgacggg tttaaaatag caaaactctg aggccacaca atagcttggg 300
cttatatggg ctcctgtggg ggaaggggga gcacggaggg ggccggggcc gctgctgcca 360
aaatagcagc tcacaagtgt tgcattcctc tctgggcgcc gggcacattc ctgctggctc 420
tgcccgcccc ggggtgggcg ccggggggac cttaaagcct ctgcccccca aggagccctt 480
cccagacagc cgccggcacc caccgctccg tgggacgatc cccgaagctc tagagcttta 540
ttgcggtagt ttatcacagt taaattgcta acgcagtcag tgcttctgac acaacagtct 600
cgaacttaag ctgcagaagt tggtcgtgag gcactgggca ggtaagtatc aaggttacaa 660
gacaggttta aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt 720
tctgataggc acctattggt cttactgaca tccactttgc ctttctctcc acaggtgtcc 780
actcccagtt caattacagc tcttaaggct agagtactta atacgactca ctatag 836
Claims (6)
1. A method of culturing cardiomyocytes in vitro for non-disease diagnostic and therapeutic purposes, comprising adding naphazoline or a pharmaceutically acceptable salt thereof to a medium containing cardiomyocytes.
2. The method according to claim 1, characterized in that: the final concentration of the naphazoline or the pharmaceutically acceptable salt thereof in the culture medium is 0.5-2 mu mol/L.
3. The method according to claim 1 or 2, characterized in that: the cardiomyocyte is a human or mammalian cardiomyocyte.
4. A method for promoting cardiomyocyte proliferation in vitro for non-disease diagnostic and therapeutic purposes, comprising treating cardiomyocytes with naphazoline or a pharmaceutically acceptable salt thereof.
5. The method according to claim 4, wherein: the concentration of the naphazoline or the pharmaceutically acceptable salt thereof in the treatment system is 0.5-2 mu mol/L.
6. The method according to claim 4 or 5, characterized in that: the cardiomyocyte is a human or mammalian cardiomyocyte.
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| Yuan Huang 等.An α1A-Adrenergic–Extracellular Signal-Regulated Kinase Survival Signaling Pathway in Cardiac Myocytes.《Circulation》.2007,第115卷(第6期),第763-772页. * |
| 孟繁浩等.《药物化学》.中国医药科技出版社,2016,第150-151页. * |
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